植物学报 ›› 2002, Vol. 19 ›› Issue (04): 444-451.

• 研究论文 • 上一篇    下一篇

向日葵品种鉴定和纯度分析的AFLP研究

刘杰 刘公社 齐冬梅 汪恩华 李芳芳   

  1. (中国科学院植物研究所 北京 100093)
  • 收稿日期:2001-07-06 修回日期:2001-08-08 出版日期:2002-07-20 发布日期:2002-07-20
  • 通讯作者: 刘公社

Study on AFLP Markers Used in Identification and Genetic Purity Test of Sunflower (Helianthus annuus L.)

LIU Jie LIU Gong-SheQI Dong-Mei WANG En-Hua LI Fang-Fang   

  1. (Institute of Botany, The Chinese Academy of Sciences, Beijing 100093)
  • Received:2001-07-06 Revised:2001-08-08 Online:2002-07-20 Published:2002-07-20
  • Contact: LIU Gong-She

摘要: 从杂交油葵A15及其亲本的1/2粒干种子中提取基因组DNA,选用17对引物组合进 行AFLP分析,构建了它们的指纹图谱。17对引物在A系与R系当中共扩增出1125条扩增产物,其中144条带表现出多态性,平均每对引物扩增66条带,不同引物组合产生的DNA片段数目在50~70之间,大小分布于100bp~500bp,多态性比率为12.8%。从中筛选出的2对引物E_AAC/M_CTC和E_ACG/M_CTG可将亲本和子代区分开:引物对E_AAC/M_CTC在A系中扩增出440bp、190bp、160bp 3条特征谱带,在R系中扩增出380bp、350bp、225bp、180bp 4条特征谱带,E_ACG/M_CTG在A系中扩增出了2条特征带480bp和265bp,在R系中扩增出490bp、220bp、205bp、125bp 4条特征谱带,且上述谱带均在子代中出现。用引物组合E_ACG/M_CTG对A15、双亲以及与A15外型十分相似的10个常用油葵杂交种进行AFLP分析,不仅表现出良好的多态性,并能够清楚地将它们加以区分。以其对50粒A15杂交种子进行纯度鉴定,得到与大田纯度检测一致的结果,说明使用AFLP标记检测油用向日葵的品种和纯度是可行的。对现行种子纯度和品种鉴定的方法进行了讨论。

Abstract: The genomic DNAs extracted from 1/2 mature seed of oilseed sunflower hybrid A15 and its parents were analyzed using AFLP (Amplified fragment length polymorphisms) to generate DNA fingerprints. A total of 1125 AFLP markers were produce with 17 primer combinations to screen A_line and R_line, ranging from 100~500 base pairs (bp), and the polymorphism rate was about 12.8%, the average bands amplified by each primer combination were 66. Primer combination E_AAC/M_CTC, could generate characteristic bands: 440 bp、190 bp、160 bp in A_line and 380 bp、350 bp、225 bp、180 bp in R_line, whereas E_ACG/M_CTG could generate characteristic bands: 480 bp, 265 bp in A_line and 490 bp、220 bp、205 bp and 125 bp in R_line. All these characteristic bands screened were found in A15 hybrids. These bands c an be used as AFLP markers to identify different varieties in oilseed sunflower and the purity of A15 hybrid seed. Several assess methods of identification on varieties and purity of hybrid were discussed in this paper.